The structures of the lipooligosaccharide and capsule polysaccharide of <i>Campylobacter jejuni</i> genome sequenced strain NCTC 11168

Michael, Frank St.; Szymanski, Christine M.; Li, Jianjun; Chan, Kalok; Khieu, Nam Huan; Larocque, Suzon; Wakarchuk, Warren W.; Brisson, Jean‐Robert; Monteiro, Mário A.

doi:10.1046/j.1432-1033.2002.03201.x

Cited by 152 publications

(192 citation statements)

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“…by phospho-form transferase A (39) have previously been observed, and pEtN modification of LOS has also been documented in a number of C. jejuni strains (37,38). In N. gonorrhoeae there is recognition of the interplay between glycosylation and pEtN modification (39,40), with overlapping site occupation observed between the O-linked glycosylation system and phospho-form transferase A -mediated phospho-form modifications (59).…”

Section: Discussionmentioning

confidence: 87%

“…In contrast to the apparently conserved nature of the N-glycan, other carbohydrate structures such as capsule polysaccharide and LOS are subject to phase variation and can also be modified by phosphate-containing moieties, including phosphoramidate (36 -37) and phosphoethanolamine (pEtN (19,38)). The addition of similar moieties to protein substrates has been documented in other pathogens, most notably the decoration of Neisseria gonorrhoeae pilin with pEtN, phosphocholine (39,40), and phosphoglycerol (41,42).…”

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confidence: 99%

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Modification of the Campylobacter jejuni N-Linked Glycan by EptC Protein-mediated Addition of Phosphoethanolamine

Scott

Nothaft

Edwards

et al. 2012

Journal of Biological Chemistry

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Section: Discussionmentioning

confidence: 87%

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confidence: 99%

Modification of the Campylobacter jejuni N-Linked Glycan by EptC Protein-mediated Addition of Phosphoethanolamine

Scott

Nothaft

Edwards

et al. 2012

Journal of Biological Chemistry

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“…NIH-PA Author Manuscript NIH-PA Author Manuscript (7)(8)(9)(10). The genes coding for enzymes that synthesize these glycan moieties have been found in clusters throughout the C. jejuni genome.…”

Section: Nih-pa Author Manuscriptmentioning

confidence: 99%

“…Pseudaminic acid, the major modification of flagellin, is synthesized via the UDP-GlcNAc dehydratase PseB pathway (30,18). GalNAc, a major component in the biosynthesis of glycoprotein N-linked heptasaccharide, lipooligosaccharide (LOS), and capsule (7,8,10), is synthesized intracellularly by the C4 epimerase Gne (31). Table 3 lists the kinetic parameters of PseB and Gne derived from other studies, as well as GST-PglF from this study.…”

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confidence: 99%

In Vitro Biosynthesis of UDP-N,N‘-Diacetylbacillosamine by Enzymes of the Campylobacter jejuni General Protein Glycosylation System

et al. 2006

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In Campylobacter jejuni the sugar 2,4-diacetamido-2,4,6-trideoxy-α-D-glucopyranose, termed N,N ′-diacetylbacillosamine (Bac2,4diNAc), is the first carbohydrate in the glycoprotein N-linked heptasaccharide. Starting with uridine diphosphate-N-acetylglucosamine (UDP-GlcNAc) two enzymes of the general protein glycosylation (Pgl) pathway in C. jejuni (PglF and PglE) have been recently shown to modify this sugar nucleotide to form UDP-2-acetamido-4-amino-2,4,6-trideoxy-α-D-glycopyranose (UDP-4-amino-sugar) [Schoenhofen, I. C., et al. (2006) J Biol Chem 281, 723-732]. PglD has been proposed to catalyze the final step in N,N′-diacetylbacillosamine synthesis by N-acetylation of the UDP-4-amino-sugar at the C4 position. We have cloned, overexpressed and purified PglD from the pgl locus of C. jejuni NCTC 11168 and identified it as the acetyltransferase that modifies the UDP-4-amino-sugar to form UDP-N,N′-diacetylbacillosamine, utilizing acetyl coenzyme A as the acetyl group donor. The UDP-N,N′-diacetylbacillosamine product was purified from the reaction by reverse phase C18 HPLC and the structure determined by NMR analysis. Additionally, the full-length PglF was overexpressed and purified in the presence of detergent as a GST-fusion protein allowing for derivation of kinetic parameters. We found that the UDP-4-aminosugar was readily synthesized from UDP-GlcNAc in a coupled reaction using PglF and PglE. We also demonstrate the in vitro biosynthesis of the complete heptasaccharide lipid-linked donor by coupling the action of eight enzymes (PglF, PglE, PglD, PglC, PglA, PglJ, PglH, and PglI) in the Pgl pathway in a single reaction vessel.Campylobacter jejuni is the Gram-negative enteropathogen identified as the primary cause of gastroenteritis in humans and the most frequent infection to precede the peripheral neuropathy Guillain-Barré syndrome (1-3). Infection of humans generally occurs by ingestion of contaminated livestock or water. Although the mechanism of infection is not clearly understood, production of glycolipids and glycoproteins by the pathogen have been found to influence cell motility, host-cell interactions, and competence for DNA uptake (4-6). As resistance to antimicrobial agents rises, the potential for development of novel therapeutics against enzymes that produce glycoconjugates has intensified efforts targeted at the characterization of their biosynthetic pathways. In C. jejuni four major glycan structures have been identified: lipooligosaccharide (LOS), capsule, O-linked glycan, and N-linked glycan *To whom correspondence should be addressed: Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, MA 02139. E-mail: imper@mit.edu, (v) 617-253-1838, (f) NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript (7)(8)(9)(10). The genes coding for enzymes that synthesize these glycan moieties have been found in clusters throughout the C. jejuni genome. The respective biochemical functions of the gene products have been assigned on the b...

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“…B, the Pgl heptasaccharide (7). C, the capsule (6). In all three diagrams, the GalNAc, Gal, GalfNAc residues appear in boldface type.…”

Section: Fig 1 the Structures Of The Glyconjugates Of C Jejuni Nctmentioning

confidence: 99%

A Single Bifunctional UDP-GlcNAc/Glc 4-Epimerase Supports the Synthesis of Three Cell Surface Glycoconjugates in Campylobacter jejuni

Bernatchez

Szymanski

Ishiyama

et al. 2005

Journal of Biological Chemistry

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The major cell-surface carbohydrates (lipooligosaccharide, capsule, and glycoprotein N-linked heptasaccharide) of Campylobacter jejuni NCTC 11168 contain Gal and/or GalNAc residues. GalE is the sole annotated UDP-glucose 4-epimerase in this bacterium. The presence of GalNAc residues in these carbohydrates suggested that GalE might be a UDP-GlcNAc 4-epimerase. GalE was shown to epimerize UDP-Glc and UDP-GlcNAc in coupled assays with C. jejuni glycosyltransferases and in sugar nucleotide epimerization equilibria studies. Thus, GalE possesses UDP-GlcNAc 4-epimerase activity and was renamed Gne. The K m(app) values of a purified MalE-Gne fusion protein for UDP-GlcNAc and UDP-GalNAc are 1087 and 1070 M, whereas those for UDP-Glc and UDP-Gal are 780 and 784 M. The k cat and k cat /K m(app) values were three to four times higher for UDP-GalNAc and UDP-Gal than for UDP-GlcNAc and UDP-Glc. The comparison of the kinetic parameters of MalE-Gne to those of other characterized bacterial UDPGlcNAc 4-epimerases indicated that Gne is a bifunctional UDP-GlcNAc/Glc 4-epimerase. The UDP sugarbinding site of Gne was modeled by using the structure of the UDP-GlcNAc 4-epimerase WbpP from Pseudomonas aeruginosa. Small differences were noted, and these may explain the bifunctional character of the C. jejuni Gne. In a gne mutant of C. jejuni, the lipooligosaccharide was shown by capillary electrophoresis-mass spectrometry to be truncated by at least five sugars. Furthermore, both the glycoprotein N-linked heptasaccharide and capsule were no longer detectable by high resolution magic angle spinning NMR. These data indicate that Gne is the enzyme providing Gal and GalNAc residues with the synthesis of all three cell-surface carbohydrates in C. jejuni NCTC 11168.

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The structures of the lipooligosaccharide and capsule polysaccharide of Campylobacter jejuni genome sequenced strain NCTC 11168

Cited by 152 publications

References 60 publications

Modification of the Campylobacter jejuni N-Linked Glycan by EptC Protein-mediated Addition of Phosphoethanolamine

Modification of the Campylobacter jejuni N-Linked Glycan by EptC Protein-mediated Addition of Phosphoethanolamine

In Vitro Biosynthesis of UDP-N,N‘-Diacetylbacillosamine by Enzymes of the Campylobacter jejuni General Protein Glycosylation System

A Single Bifunctional UDP-GlcNAc/Glc 4-Epimerase Supports the Synthesis of Three Cell Surface Glycoconjugates in Campylobacter jejuni

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